Antibodies to human il-1beta

ABSTRACT

An IL-1β binding molecule, in particular an antibody to human IL-1β, especially a human antibody to human IL-1β is provided, wherein the CDRs of the heavy and light chains have amino acid sequences as defined, for use in the treatment of an IL-1 mediated disease or disorder, e.g. osteoarthritis, osteoporosis and other inflammatory arthritides.

This is a divisional of application Ser. No. 11/484,472 filed on Jul.11, 2006, which is a continuation of application Ser. No. 10/181,324filed on Jul. 16, 2002, which is a 371 of Application No.PCT/EP01/00591, filed on Jan. 19, 2001, which claims priority to GB0001448.0. filed Jan. 21, 2000, the entire disclosures of which arehereby incorporated by reference.

This invention relates to antibodies to human interleukin 1 beta (IL-1β)and to the use of such antibodies for the treatment of IL-1 mediateddiseases and disorders.

Interleukin 1 (IL-1) is an activity produced by cells of the immunesystem which acts as a mediator of the acute phase inflammatoryresponse. Inappropriate or excessive production of IL-1, in particularIL-1β, is associated with the pathology of various diseases anddisorders, such as septicemia, septic or endotoxic shock, allergies,asthma, bone loss, ischemia, stroke, rheumatoid arthritis and otherinflammatory disorders. Antibodies to IL-1β have been proposed for usein the treatment of IL-1 mediated diseases and disorders; see forinstance, WO 95/01997 and the discussion in the introduction thereof.

We have now prepared improved antibodies to human IL-1β for use in thetreatment of IL-1 mediated diseases and disorders.

Accordingly the invention provides an IL-1β binding molecule whichcomprises an antigen binding site comprising at least one immunoglobulinheavy chain variable domain (V_(H)) which comprises in sequencehypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the aminoacid sequence Ser-TyT-Trp-Ile-Gly (SEQ ID NO:5), said CDR having theamino acid sequenceIle-Ile-TyT-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly (SEQID NO:6), and said CDR3 having the amino acid sequenceTyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile (SEQ ID NO:7); and directequivalents thereof.

In a first aspect the invention provides a single domain IL-1β bindingmolecule comprising an isolated immunoglobulin heavy chain comprising aheavy chain variable domain (V_(H)) as defined above.

In a second aspect the invention also provides an IL-1β binding moleculecomprising both heavy (V_(H)) and light chain (V_(L)) variable domainsin which said IL-1β binding molecule comprises at least one antigenbinding site comprising:

-   -   a) an immunoglobulin heavy chain variable domain (V_(H)) which        comprises in sequence hypervariable regions CDR1, CDR and CDR3,        said CDR1 having the amino acid sequence Ser-Tyr-Trp-Ile-Gly        (SEQ ID NO:5), said CDR2 having the amino acid sequence        Ile-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly        (SEQ ID NO:6), and said CDR3 having the amino acid sequence        Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile (SEQ ID NO:7), and    -   b) an immunoglobulin light chain variable domain (V_(L)) which        comprises a CDR3′ hypervariable region having the amino acid        sequence Gln-Gln-Arg-Ser-Asn-Trp-Met-Phe-Pro (SEQ ID NO:10);        and direct equivalents thereof.

In particular embodiments of the second aspect the invention provides anIL-1β binding molecule comprising both heavy (V_(H)) and light (V_(L))chain variable domains in which said IL-1β binding molecule comprises atleast one antigen binding site comprising:

-   -   a) an immunoglobulin heavy chain variable domain (V_(H)) which        comprises in sequence hypervariable regions CDR1, CDR2 and CDR3,        said CDR1 having the amino acid sequence Ser-Tyr-Trp-Ile-Gly        (SEQ ID NO:5), said CDR having the amino acid sequence        Ile-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly        (SEQ ID NO:6), and said CDR3 having the amino acid sequence        Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile (SEQ ID NO:7), and    -   b) an immunoglobulin light chain variable domain (V_(L)) which        comprises in sequence hypervariable regions CDR1′, CDR2′ and        CDR3′, said CDR1′ having the amino acid sequence        Arg-Ala-Ser-Gln-Ser-Val-Ser-Ser-Tyr-Leu Ala (SEQ ID NO:8), said        CDR2′ having the amino acid sequence Asp-Ala-Ser-Asn-Arg-Ala-Thr        (SEQ ID NO:9), and said CDR3′ having the amino acid sequence        Gln-Gln-Arg-Ser-Asn-Trp-Met-Phe-Pro (SEQ ID NO:10);        and direct equivalents thereof.

Unless otherwise indicated, any polypeptide chain is herein described ashaving an amino acid sequence starting at the N-terminal extremity andending at the C-terminal extremity. When the antigen binding sitecomprises both the V_(H) and V_(L) domains, these may be located on thesame polypeptide molecule or, preferably, each domain may be on adifferent chain, the V_(H) domain being part of an immunoglobulin heavychain or fragment thereof and the V_(L) being part of an immunoglobulinlight chain or fragment thereof.

By “IL-1β binding molecule” is meant any molecule capable of binding tothe IL-1β antigen either alone or associated with other molecules. Thebinding reaction may be shown by standard methods (qualitative assays)including, for example, a bioassay for determining the inhibition ofIL-1β binding to its receptor or any kind of binding assays, withreference to a negative control test in which an antibody of unrelatedspecificity but of the same isotype, e.g. an anti-CD25 antibody, isused. Advantageously, the binding of the IL-1β binding molecules of theinvention to IL-1β may be shown in a competitive binding assay.

Examples of antigen binding molecules include antibodies as produced byB-cells or hybridomas and chimeric, CDR-grafted or human antibodies orany fragment thereof, e.g. F(ab′)₂ and Fab fragments, as well as singlechain or single domain antibodies.

A single chain antibody consists of the variable domains of the heavyand light chains of an antibody covalently bound by a peptide linkerusually consisting of from 10 to 30 amino acids, preferably from 15 to25 amino acids. Therefore, such a structure does not include theconstant part of the heavy and light chains and it is believed that thesmall peptide spacer should be less antigenic than a whole constantpart. By “chimeric antibody” is meant an antibody in which the constantregions of heavy or light chains or both are of human origin while thevariable domains of both heavy and light chains are of non-human (e.g.murine) origin or of human origin but derived from a different humanantibody. By “CDR-grafted antibody” is meant an antibody in which thehypervariable regions (CDRs) are derived from a donor antibody, such asa non-human (e.g. murine) antibody or a different human antibody, whileall or substantially all the other parts of the immunoglobulin e.g. theconstant regions and the highly conserved parts of the variable domains,i.e. the framework regions, are derived from an acceptor antibody, e.g.an antibody of human origin. A CDR-grafted antibody may however containa few amino acids of the donor sequence in the framework regions, forinstance in the parts of the framework regions adjacent to thehypervariable regions. By “human antibody” is meant an antibody in whichthe constant and variable regions of both the heavy and light chains areall of human origin, or substantially identical to sequences of humanorigin, not necessarily from the same antibody and includes antibodiesproduced by mice in which the murine immunoglobulin variable andconstant part genes have been replaced by their human counterparts, e.g.as described in general terms in EP 0546073 B1, U.S. Pat. No. 5,545,806,U.S. Pat. No. 5,569,825, U.S. Pat. No. 5,625,126, U.S. Pat. No.5,633,425, U.S. Pat. No. 5,661,016, U.S. Pat. No. 5,770,429, EP 0438474B1 and EP 0463151 B1.

Particularly preferred IL-1β binding molecules of the invention arehuman antibodies especially the AAL 160 antibody as hereinafterdescribed in the Examples.

Thus in preferred chimeric antibodies the variable domains of both heavyand light chains are of human origin, for instance those of the AAL 160antibody which are shown in SEQ ID NO:1 and SEQ ID NO:2. Nucleic acidsequences for the V_(L) and V_(H) chains are set forth in SEQ ID NO:3and SEQ ID NO:4, respectively. The constant region domains preferablyalso comprise suitable human constant region domains, for instance asdescribed in “Sequences of Proteins of Immunological Interest”, Kabat E.A. et al, US Department of Health and Human Services, Public HealthService, National Institute of Health

Hypervariable regions may be associated with any kind of frameworkregions, though preferably are of human origin. Suitable frameworkregions are described in Kabat E. A. et al, ibid. The preferred heavychain framework is a human heavy chain framework, for instance that ofthe AAL 160 antibody which is shown in SEQ ID NO:1. It consists insequence of FR1, FR2, FR3 and FR4 regions. In a similar manner, SEQ IDNO:2 shows the preferred AAL 160 light chain framework which consists,in sequence, of FR1′, FR2′, FR3′ and FR4′ regions.

Accordingly, the invention also provides an IL-1β binding molecule whichcomprises at least one antigen binding site comprising either a firstdomain having an amino acid sequence substantially identical to thatshown in SEQ ID NO:1 starting with amino acid at position 1 and endingwith amino acid at position 118 or a first domain as described above anda second domain having an amino acid sequence substantially identical tothat shown in SEQ ID NO:2, starting with amino acid at position 1 andending with amino acid at position 107.

Monoclonal antibodies raised against a protein naturally found in allhumans are typically developed in a nonhuman system e.g. in mice. As adirect consequence of this, a xenogenic antibody as produced by ahybridoma, when administered to humans, elicits an undesirable immuneresponse which is predominantly mediated by the constant part of thexenogenic immunoglobulin. This clearly limits the use of such antibodiesas they cannot be administered over a prolonged period of time.Therefore it is particularly preferred to use single chain, singledomain, chimeric, CDR-grafted, or especially human antibodies which arenot likely to elicit a substantial allogenic response when administeredto humans.

In view of the foregoing, a more preferred IL-1β binding molecule of theinvention is selected from a human anti IL-1β antibody which comprisesat least

-   -   a) an immunoglobulin heavy chain or fragment thereof which        comprises (i) a variable domain comprising in sequence the        hypervariable regions CDR1, CDR and CDR3 and (ii) the constant        part or fragment thereof of a human heavy chain; said CDR1        having the amino acid sequence Ser-Tyr-Trp-Ile-Gly (SEQ ID        NO:5), said CDR having the amino acid sequence        Ile-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly        (SEQ ID NO:6), and said CDR3 having the amino acid sequence        Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile (SEQ ID NO:7) and    -   b) an immunoglobulin light chain or fragment thereof which        comprises (i) a variable domain comprising the CDR3′        hypervariable region and optionally also the CDR1′, CDR2′        hypervariable regions and (ii) the constant part or fragment        thereof of a human light chain, said CDR1′ having the amino acid        sequence Arg-Ala-Ser-Gln-Ser-Val-Ser-Ser-Tyr-Leu Ala (SEQ ID        NO:8), said CDR2′ having the amino acid sequence        Asp-Ala-Ser-Asn-Arg-Ala-Thr (SEQ ID NO:9), and said CDR3′ having        the amino acid sequence Gln-Gln-Arg-Ser-Asn-Trp-Met-Phe-Pro (SEQ        ID NO:10); and direct equivalents thereof.

Alternatively, an IL-1β binding molecule of the invention may beselected from a single chain binding molecule which comprises an antigenbinding site comprising

-   a) a first domain comprising in sequence the hypervariable regions    CDR1, CDR2 and CDR3, said hypervariable regions having the amino    acid sequences as shown in SEQ ID NO:1,-   b) A second domain comprising the hypervariable regions CDR3′ and    optionally CDR1′ and CDR2′, said hypervariable regions having the    amino acid sequences as shown in SEQ ID NO:2 and-   c) a peptide linker which is bound either to the N-terminal    extremity of the first domain and to the C-terminal extremity of the    second domain or to the C-terminal extremity of the first domain and    to the N-terminal extremity of second domain;    and direct equivalents thereof.

As it is well known, minor changes in an amino acid sequence such asdeletion, addition or substitution of one, a few or even several aminoacids may lead to an allelic form of the original protein which hassubstantially identical properties.

Thus, by the term “direct equivalents thereof” is meant either anysingle domain IL-1 binding molecule (molecule X).

-   (i) in which the hypervariable regions CDR1, CDR2 and CDR3 taken as    a whole are at least 80% homologous, preferably at least 90%    homologous, more preferably at least 95% homologous to the    hypervariable regions as shown in SEQ ID NO:1 and,-   (ii) which is capable of inhibiting the binding of IL-1β to its    receptors substantially to the same extent as a reference molecule    having framework regions identical to those of molecule X but having    hypervariable regions CDR1, CDR2 and CDR3 identical to those shown    in SEQ ID NO:1    or any IL-1β binding molecule having at least two domains per    binding site (molecule X′)-   (i) in which the hypervariable regions CDR1, CDR2, CDR3, CDR3′ and    optionally CDR1′ and CDR2′ taken as a whole are at least 80%    homologous, preferably at least 90% homologous, more preferably at    least 95% homologous, to the hypervariable regions as shown in SEQ    ID NO:1 and 2 and-   (ii) which is capable of inhibiting the binding of IL-1β to its    receptors substantially to the same extent as a reference molecule    having framework regions and constant parts identical to molecule    X′, but having hypervariable regions CDR1, CDR2, CDR3, and CDR3′,    and optionally CDR1 ′and CDR2′, identical to those shown in SEQ ID    NO:1 and 2.

In the present description amino acid sequences are at least 80%homologous to one another if they have at least 80% identical amino acidresidues in a like position when the sequence are aligned optimally,gaps or insertions in the amino acid sequences being counted asnon-identical residues.

The inhibition of the binding of IL-1β to its receptor may beconveniently tested in various assays including such assays aredescribed hereinafter in the text. The IL-1β receptor used is preferablythe IL-1β type 1 receptor. By the term “to the same extent” is meantthat the reference and the equivalent molecules exhibit, on astatistical basis, essentially identical IL-1β binding inhibition curvesin one of the assays referred to above.

For example, the assay used may be an assay of competitive inhibition ofbinding of IL-1β by soluble IL-1 receptors and the IL-1β bindingmolecules of the invention.

Most preferably, the human IL-1β antibody comprises at least

-   -   a) one heavy chain which comprises a variable domain having an        amino acid sequence substantially identical to that shown in SEQ        ID NO:1 starting with the amino acid at position 1 and ending        with the amino acid at position 118 and the constant part of a        human heavy chain; and    -   b) one light chain which comprises a variable domain having an        amino acid sequence substantially identical to that shown in SEQ        ID NO:2 starting with the amino acid at position 1 and ending        with the amino acid at position 107 and the constant part of a        human light chain.

The constant part of a human heavy chain may be of the γ₁, γ₂, γ₃, γ₄,μ, α₁, α₂, δ or ε type, preferably of the γ type, more preferably of theγ₁ type, whereas the constant part of a human light chain may be of theκ or λ type (which includes the λ₁, λ₂ and λ₃ subtypes) but ispreferably of the κ type. The amino acid sequences of all these constantparts are given in Kabat et al ibid.

An IL-1β binding molecule of the invention may be produced byrecombinant DNA techniques. In view of this, one or more DNA moleculesencoding the binding molecule must be constructed, placed underappropriate control sequences and transferred into a suitable hostorganism for expression.

In a very general manner, there are accordingly provided

-   (i) DNA molecules encoding a single domain IL-1β binding molecule,    of the invention, a single chain IL-1β binding molecule of the    invention, a heavy or light chain or fragments thereof of a IL-1β    binding molecule of the invention and-   (ii) the use of the DNA molecules of the invention for the    production of a IL-1β binding molecule of the invention by    recombinant means.

The present state of the art is such that the skilled worker in the artis able to synthesize the DNA molecules of the invention given theinformation provided herein i.e. the amino acid sequences of thehypervariable regions and the DNA sequences coding for them. A methodfor constructing a variable domain gene is for example described in EPA239 400 and may be briefly summarized as follows: A gene encoding avariable domain of a MAb of whatever specificity is cloned. The DNAsegments encoding the framework and hypervariable regions are determinedand the DNA segments encoding the hypervariable regions are removed sothat the DNA segments encoding the framework regions are fused togetherwith suitable restriction sites at the junctions. The restriction sitesmay be generated at the appropriate positions by mutagenesis of the DNAmolecule by standard procedures. Double stranded synthetic CDR cassettesare prepared by DNA synthesis according to the sequences given in SEQ IDNO:1 or 2. These cassettes are provided with sticky ends so that theycan be ligated at the junctions of the framework

Furthermore, it is not necessary to have access to the mRNA from aproducing hybridoma cell line in order to obtain a DNA construct codingfor the IL-1β binding molecules of the invention. Thus PCT applicationWO 90/107861 gives full instructions for the production of an antibodyby recombinant DNA techniques given only written information as to thenucleotide sequence of the gene. The method comprises the synthesis of anumber of oligonucleotides, their amplification by the PCR method, andtheir splicing to give the desired DNA sequence.

Expression vectors comprising a suitable promoter or genes encodingheavy and light chain constant parts are publicly available. Thus, oncea DNA molecule of the invention is prepared it may be convenientlytransferred in an appropriate expression vector. DNA molecules encodingsingle chain antibodies may also be prepared by standard methods, forexample, as described in WO 88/1649.

In view of the foregoing no hybridoma or cell line deposit is necessaryto comply with the criteria of sufficiency of description.

In a particular embodiment the invention includes first and second DNAconstructs for the production of an IL-1β binding molecule as describedbelow:

The first DNA construct encodes a heavy chain or fragment thereof andcomprises

-   -   a) a first part which encodes a variable domain comprising        alternatively framework and hypervariable regions, said        hypervariable regions being in sequence CDR1, CDR2 and CDR3 the        amino acid sequences of which are shown in SEQ ID NO:1; this        first part starting with a codon encoding the first amino acid        of the variable domain and ending with a codon encoding the last        amino acid of the variable domain, and    -   b) a second part encoding a heavy chain constant part or        fragment thereof which starts with a codon encoding the first        amino acid of the constant part of the heavy chain and ends with        a codon encoding the last amino acid of the constant part or        fragment thereof, followed by a stop codon.

Preferably, this first part encodes a variable domain having an aminoacid sequence substantially identical to the amino acid sequence asshown in SEQ ID NO:1 starting with the amino acid at position 1 andending with the amino acid at position 118. More preferably the firstpart has the nucleotide sequence as shown in SEQ ID NO:1 starting withthe nucleotide at position 1 and ending with the nucleotide at position354. Also preferably, the second part encodes the constant part of ahuman heavy chain, more preferably the constant part of the human γ1chain. This second part may be a DNA fragment of genomic origin(comprising introns) or a cDNA fragment (without introns).

The second DNA construct encodes a light chain or fragment thereof andcomprises

-   -   a) a first part which encodes a variable domain comprising        alternatively framework and hypervariable regions; said        hypervariable regions being CDR3′ and optionally CDR1′ and        CDR2′, the amino acid sequences of which are shown in SEQ ID        NO:2; this first part starting with a codon encoding the first        amino acid of the variable domain and ending with a codon        encoding the last amino acid of the variable domain, and    -   b) a second part encoding a light chain constant part or        fragment thereof which starts with a codon encoding the first        amino acid of the constant part of the light chain and ends with        a codon encoding the last amino acid of the constant part or        fragment thereof followed by a stop codon.

Preferably, this first part encodes a variable domain having an aminoacid sequence substantially identical to the amino acid sequence asshown in SEQ ID NO:2 starting with the amino acid at position 1 andending with the amino acid at position 107. More preferably, the firstpart has the nucleotide sequence as shown in SEQ ID NO:2 starting withthe nucleotide at position 1 and ending with the nucleotide at position321. Also preferably the second part encodes the constant part of ahuman light chain, more preferably the constant part of the human κchain.

The invention also includes IL-1β binding molecules in which one or moreof the residues of CDR1, CDR2, CDR3, CDR1′, CDR2′ or CDR3′ are changedfrom the residues shown in SEQ ID NO:1 and SEQ ID NO:2 for instance bymutation e.g. site directed mutagenesis of the corresponding DNAsequences. The invention includes the DNA sequences coding for suchchanged IL-1β binding molecules. In particular the invention includesIL-1β binding molecules in which one or more residues of CDR1′ or CDR2′have been changed from the residues shown in SEQ ID NO:2.

In the first and second DNA constructs, the first and second parts maybe separated by an intron, and, an enhancer may be conveniently locatedin the intron between the first and second parts. The presence of suchan enhancer which is transcribed but not translated, may assist inefficient transcription. In particular embodiments the first and secondDNA constructs comprise the enhancer of a heavy chain geneadvantageously of human origin.

Each of the DNA constructs are placed under the control of suitablecontrol sequences, in particular under the control of a suitablepromoter. Any kind of promoter may be used, provided that it is adaptedto the host organism in which the DNA constructs will be transferred forexpression. However, if expression is to take place in a mammalian cell,it is particularly preferred to use the promoter of an immunoglobulingene, or a cytomegalovirus (CMV) promoter, e.g. a human CMV promoter.

The desired antibody may be produced in a cell culture or in atransgenic animal. A suitable transgenic animal may be obtainedaccording to standard methods which include micro injecting into eggsthe first and second DNA constructs placed under suitable controlsequences transferring the so prepared eggs into appropriatepseudo-pregnant females and selecting a descendant expressing thedesired antibody.

When the antibody chains are produced in a cell culture, the DNAconstructs must first be inserted into either a single expression vectoror into two separate but compatible expression vectors, the latterpossibility being preferred.

Accordingly, the invention also provides an expression vector able toreplicate in a prokaryotic or eukaryotic cell line which comprises atleast one of the DNA constructs above described.

Each expression vector containing a DNA construct is then transferredinto a suitable host organism. When the DNA constructs are separatelyinserted on two expression vectors, they may be transferred separately,i.e. one type of vector per cell, or co-transferred, this latterpossibility being preferred. A suitable host organism may be abacterium, a yeast or a mammalian cell line, this latter beingpreferred. More preferably, the mammalian cell line is of lymphoidorigin, e.g. a myeloma, hybridoma or a normal immortalised B-cell, whichconveniently does not express any endogenous antibody heavy or lightchain.

For expression in mammalian cells it is preferred that the IL-1β bindingmolecule coding sequence is integrated into the host cell DNA within alocus which permits or favours high level expression of the IL-1βbinding molecule. Cells in which the IL-1β binding molecule codingsequence is integrated into such favourable loci may be identified andselected on the basis of the levels of the IL-1β binding molecule whichthey express. Any suitable selectable marker may be used for preparationof host cells containing the IL-1β binding molecule coding sequence; forinstance, a dhfr gene/methotrexate or equivalent selection system may beused. Preferred systems for expression of the IL-1β binding molecules ofthe invention include GS-based amplification selection systems, such asthose described in EP 0256055 B, EP 0323997 B and European patentapplication 89303964.4. Preferably also the vector may contain othersequences as desired to facilitate expression, processing and export ofthe expressed protein; for example, the vector may typically contain aleader sequence associated with the coding sequence.

In a further aspect of the invention there is provided a process for theproduct of an IL-1β binding molecule which comprises (i) culturing anorganism which is transformed with an expression vector as defined aboveand (ii) recovering the IL-1β binding molecule from the culture.

In accordance with the present invention it has been found that the AAL160 antibody has binding specificity for the antigenic epitope of humanIL-1β which includes the loop comprising residues, Gly 22, Pro 23, Tyr24 and Glu 25 of mature human IL-1β. (Residues, Gly 22, Pro 23, Tyr 24and Glu 25 of mature human IL-1β correspond to residues 138, 139, 140and 141 respectively of the human IL-1β precursor.) This epitope appearsto be outside the recognition site of the IL-1 receptor and it istherefore most surprising that antibodies to this epitope, e.g. theAAL160 antibody, are capable of inhibiting the binding of IL-1β to itsreceptor. Antibodies, in particular chimeric and CDR-grafted antibodiesand especially human antibodies, which have binding specificity for theantigenic epitope of mature human IL-1β which includes the loopcomprising residues, Gly 22, Pro 23, Tyr 24 and Glu 25 and which arecapable of inhibiting the binding of IL-1β to its receptor; and use ofsuch antibodies for the treatment of IL-1 mediated diseases anddisorders, are novel and are included within the scope of the presentinvention.

Thus in a further aspect the invention includes an antibody to IL-1βwhich has antigen binding specificity for an antigenic epitope of humanIL-1β which includes the loop comprising residues Gly 22, Pro 23, Tyr 24and Glu 25 of mature human IL-1β of mature human IL-1β and which iscapable of inhibiting the binding of IL-1β to its receptor.

In yet further aspects the invention includes:

-   -   i) use of an antibody to IL-1β, which has antigen binding        specificity for an antigenic epitope of mature human IL-1β which        includes the loop comprising residues Gly 22, Pro 23, Tyr 24 and        Glu 25 and which is capable of inhibiting the binding of IL-1β        to its receptor, for the treatment of an IL-1 mediated disease        or disorder;    -   ii) a method for the treatment of an IL-1 mediated disease or        disorders in a patient which comprises administering to the        patient an effective amount of an antibody to IL-1β, which has        antigen binding specificity for an antigenic epitope of mature        human IL-1β which includes the loop comprising residues Gly 22,        Pro 23, Tyr 24 and Glu 25 and which is capable of inhibiting the        binding of IL-1β to its receptor;    -   iii) a pharmaceutical composition comprising an antibody to        IL-1β, which has antigen binding specificity for an antigenic        epitope of mature human IL-1β which includes the loop comprising        residues Gly 22, Pro 23, Tyr 24 and Glu 25 and which is capable        of inhibiting the binding of IL-1β to its receptor, in        combination with a pharmaceutically acceptable excipient,        diluent or carrier; and    -   iv) use of an antibody to IL-1β, which has antigen binding        specificity for an antigenic epitope of mature human IL-1β which        includes the loop comprising residues Gly 22, Pro 23, Tyr 24 and        Glu 25 and which is capable of inhibiting the binding of IL-1β        to its receptor, for the preparation of a medicament for the        treatment of an IL-1 mediated disease or disorder.

For the purposes of the present description an antibody is “capable ofinhibiting the binding of IL-1β” if the antibody is capable ofinhibiting the binding of IL-1β to its receptor substantially to thesame extent as the AAL160 antibody, wherein “to the same extent” hasmeaning as defined above.

In the present description the phrase “IL-1 mediated disease”encompasses all diseases and medical conditions in which IL-1 plays arole, whether directly or indirectly, in the disease or medicalcondition, including the causation, development, progress, persistenceor pathology of the disease or condition.

In the present description the terms “treatment” or “treat” refer toboth prophylactic or preventative treatment as well as curative ordisease modifying treatment, including treatment of patient at risk ofcontracting the disease or suspected to have contracted the disease aswell as patients who are ill or have been diagnosed as suffering from adisease or medical condition, and includes suppression of clinicalrelapse.

Antibodies which have binding specificity for the antigenic epitope ofmature human IL-1, which includes the loop comprising residues Gly 22,Pro 23, Tyr 24 and Glu 25 and which are capable of inhibiting thebinding of IL-1β to its receptor are hereinafter referred to asAntibodies of the Invention. Preferably Antibodies of the Invention areantibodies which have binding specificity for this epitope of humanIL-1β when the human IL-1β is under native, e.g. normal physiologicalconditions, not under denatured conditions, e.g. not in the presence ofa denaturing agent such as SDS. Antibodies of the Invention maycross-react with non-human IL-1βs, which have antigenic epitopes whichinclude Gly at residue 22, Pro at residue 23, Tyr at residue 24 and Gluat residue 25 and which are closely similar to the corresponding humanepitope. For example, Antibodies of the Invention may cross-react withprimate IL-1βs, such as rhesus monkey, cynomolgus monkey IL-1 ormarmoset monkey IL-1.

Preferably the Antibodies of the Invention are IL-1β binding moleculesaccording to the first and second aspects of the invention.Advantageously the Antibodies of the Invention are human antibodies,most preferably the AAL160 antibody or direct equivalent thereof.

The Antibodies of the Invention block the effects of IL-1β on its targetcells and thus are indicated for use in the treatment of IL-1 mediateddiseases and disorders. These and other pharmacological activities ofthe Antibodies of the Invention may be demonstrated in standard testmethods for example as described below:

1. Neutralization of Human IL-1β-Mediated Activation of the IL-8Promoter

The potential to neutralize IL-1β-dependent cellular signaling isdetermined in a reporter gene assay.

The human melanoma cell line G361 is stably transfected with aluciferase reporter gene construct based on the human IL-8 promoter.Reporter gene expression and activity is dependent on IL-1β or TNFα inthis cell line. Cells are stimulated with 300 pg/ml of recombinant humanIL-1β or the equivalent of 100 pg/ml in conditioned medium in thepresence of various concentrations of Antibody of the Invention or IL-1receptor antagonist ranging between 6 and 18,000 pM. The chimericantibody SIMULECT® (basiliximab) is used as a matched isotype control.Luciferase activity is quantified in a chemiluminescence assay.Antibodies of the Invention typically have IC₅₀ of about 1 nM (e.g. fromabout 0.2 to about 5 nM) when tested in this assay.

2. Neutralization of IL-1β Dependent Production of PGE₂ andInterleukin-6 by Primary Human Fibroblasts

The production of PGE₂ and IL-6 in primary human dermal fibroblasts isdependent on IL-1β. TNF-α alone cannot efficiently induce theseinflammatory mediators, but synergizes with IL-1. Primary dermalfibroblasts are used as a surrogate model for IL-1-induced cellularactivation.

Primary human fibroblasts are stimulated with recombinant IL-1β orconditioned medium obtained from LPS-stimulated human PBMCs in thepresence of various concentrations of Antibody of the Invention orIL-1RA ranging from 6 to 18,000 pM. The chimeric anti-CD25 antibodySIMULECT® (basiliximab) is used as a matched isotype control.Supernatant is taken after 16 h stimulation and assayed for IL-6 byELISA or PGE₂ by RIA. Antibodies of the Invention typically have IC₅₀sfor inhibition of IL-6 production of about 1 nM or less (e.g. from about0.1 to about 1 nM) and for inhibition of PGE₂ production of about 1 nM(e.g. from about 0.1 to about 1 nM) when tested as above.

As indicated in the above assays Antibodies of the Invention potentlyblock the effects of IL-1β. Accordingly, the Antibodies of the Inventionhave pharmaceutical utility as follows:

Antibodies of the Invention are useful for the prophylaxis and treatmentof IL-1 mediated diseases or medical conditions, e.g. inflammatoryconditions, allergies and allergic conditions, hypersensitivityreactions, autoimmune diseases, severe infections, and organ or tissuetransplant rejection.

For example, Antibodies of the Invention may be use for the treatment ofrecipients of heart, lung, combined heart-lung, liver, kidney,pancreatic, skin or corneal transplants and for the prevention ofgraft-versus-host disease, such as following bone marrow transplant.

Antibodies of the Invention are particularly useful for the treatment,prevention, or amelioration of autoimmune disease and of inflammatoryconditions, in particular inflammatory conditions with an aetiologyincluding an autoimmune component such as arthritis (for examplerheumatoid arthritis, arthritis chronica progrediente and arthritisdeformans) and rheumatic diseases, including inflammatory conditions andrheumatic diseases involving bone loss, inflammatory pain,hypersensitivity (including both airways hypersensitivity and dermalhypersensitivity) and allergies. Specific auto-immune diseases for whichAntibodies of the Invention may be employed include autoimmunehaematological disorders (including e.g. hemolytic anaemia, aplasticanaemia, pure red cell anaemia and idiopathic thrombocytopenia),systemic lupus erythematosus, polychondritis, scleredoma, Wegenergranulomatosis, dermatomyositis, chronic active hepatitis, myastheniagravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmuneinflammatory bowel disease (including e.g. ulcerative colitis, Crohn'sdisease and Irritable Bowel Syndrome), endocrine ophthalmopathy, Gravesdisease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis,juvenile diabetes (diabetes mellitus type I), uveitis (anterior andposterior), keratoconjunctivitis sicca and vernal keratoconjunctivitis,interstitial lung fibrosis, psoriatic arthritis and glomerulonephritis(with and without nephrotic syndrome, e.g. including idiopathicnephrotic syndrome or minimal change nephropathy).

Antibodies of the Invention are also useful for the treatment,prevention, or amelioration of asthma, bronchitis, pneumoconiosis,pulmonary emphysema, and other obstructive or inflammatory diseases ofthe airways

Antibodies of the Invention are useful for treating undesirable acuteand hyperacute inflammatory reactions which are mediated by IL-1 orinvolve IL-1 production, especially IL-1β, or the promotion of TNFrelease by IL-1, e.g. acute infections, for example septic shock (e.g.,endotoxic shock and adult respiratory distress syndrome), meningitis,pneumonia; and severe burns; and for the treatment of cachexia orwasting syndrome associated with morbid TNF release, consequent toinfection, cancer, or organ dysfunction, especially AIDS-relatedcachexia, e.g., associated with or consequential to HIV infection.

Antibodies of the Invention are particularly useful for treatingdiseases of bone metabolism including osteoarthritis, osteoporosis andother inflammatory arthritides, and bone loss in general, includingage-related bone loss, and in particular periodontal disease.

Antibodies of the Invention may be used for treatment of cancers, inparticular IL-1-dependent tumours.

For these indications, the appropriate dosage will, of course, varydepending upon, for example, the particular Antibody of the Invention tobe employed, the host, the mode of administration and the nature andseverity of the condition being treated. However, in prophylactic use,satisfactory results are generally indicated to be obtained at dailydosages from about 0.1 mg to about 5 mg per kilogram body weight.Antibody of the Invention is conveniently administered parenterally,intravenously, e.g. into the antecubital or other peripheral vein,intramuscularly, or subcutaneously. A prophylactic treatment typicallycomprises administering the molecule of the invention once daily to onceweekly for 2 to 4 weeks.

Pharmaceutical compositions of the invention may be manufactured inconventional manner. A composition according to the invention ispreferably provided in lyophilized form. For immediate administration itis dissolved in a suitable aqueous carrier, for example sterile waterfor injection or sterile buffered physiological saline. If it isconsidered desirable to make up a solution of larger volume foradministration by infusion, e.g. iv infusion, rather than as a bolusinjection, e.g. a sc bolus injection, it is advantageous to incorporatehuman serum albumin or the patient's own heparinised blood into thesaline at the time of formulation. The presence of an excess of suchphysiologically inert protein prevents loss of antibody by adsorptiononto the walls of the container and tubing used with the infusionsolution. If albumin is used, a suitable concentration is from 0.5 to4.5% by weight of the saline solution.

The invention is further described by way of illustration only in thefollowing Examples which refer to the accompanying Figures:

FIG. 1 which is a graph showing competitive inhibition of AAL160 bindingto IL-1β by soluble IL-1 type I and type II receptors;

FIG. 2 which is a graph showing inhibition of IL-1β-induced fever in arat model by AAL160, and

FIG. 3 which is a graph showing duration of action of AAL 160 in ratIL-1β-induced fever.

EXAMPLES

Transgenic mice engineered to express the human IgG/κ repertoire insteadof the murine immunoglobulin repertoire (Fishwild et al., 1996, NatBiotechnol., 14, 845-851) are used to generate antibodies to humanIL-1β. B cells from these mice are immortalized by standard hybridomatechnology and murine hybridoma cells are obtained which secrete thehuman IgG1/κ antibody AAL 160

Example 1 Generation of the Hybridoma and Purification of the Antibody

Genetically engineered mouse 66 (Medarex Inc. Annadale, N.J.) isimmunized with recombinant human IL-1β (50 μg) s.c. in several sites inadjuvant. The mouse is boosted five additional times with the lastinjection three days before the fusion. On the day of the fusion mouse66 is killed by CO₂ inhalation and spleen cells (4.1×10⁷) are fused by aroutine method using PEG 4000 with an equal number of PAI-O cells, amouse myeloma cell line. Fused cells are plated out in 624 wells (1ml/well) containing a feeder layer of mouse peritoneal cells (Balb Cmice), in HAT supplemented RPMI 1640, 10% heat inactivated fetal calfserum 5×10⁻⁵ M β-mercaptoethanol. Supernatants are collected and testedin ELISA and screened for IL-1β reactive monoclonal antibodies. Fivemonoclonal antibodies of the IgG/κ subclass are identified. Cloning isdone using 4×96 well microtiter plates, plating 0.5 cells per well.After two weeks wells are inspected with an inverted microscope.Supernatant is collected from wells positive for growth and productionof anti-IL-1β monoclonal antibodies is evaluated by ELISA. 1-2 L ofconditioned supernatant from four subclones of the originally identifiedhybridoma # 476 are prepared and antibodies are purified by affinitychromatography on a protein A column.

Purity and Partial Amino Acid Sequences of Heavy and Light Chain AminoAcid Sequencing

Light and heavy chains of the purified antibody AAL160 are separated bySDS-PAGE and the amino-terminal amino acids determined by Edmandegradation. The purity of the antibody used in these studies is ≧90% bysequencing. cDNA sequences coding for the heavy and light chain variabledomains are obtained by PCR amplification of cDNA obtained from mRNAfrom the cloned hybridoma cells and fully sequenced. The amino-terminalsequences of heavy and light chain variable domains and thecorresponding DNA sequences are given in SEQ ID NO:1 and SEQ ID NO:2.

The DNA sequences coding for the heavy and light chain variable domainsand the corresponding amino acid sequences of AAL160 are also give inthe accompanying sequence listing as SEQ ID NOs:3 and 4.

Construction of Expression Vectors for Heavy and Light Chain

The cloned V_(L) and V_(H) encoding sequences were amplified by PCR andinserted via appropriate restriction sites into cassette vectorsproviding the immunoglobulin promoter, the leader sequences from theRFT2 antibody (Heinrich et al. (1989) J. Immunol. 143, 3589-97), part ofthe J-segments and a splice donor site. The light chain cassettecontaining the entire V_(L) region, promoter and leader sequence forsecretion was transferred into an expression vector containing the humanCk gene, the immunoglobulin heavy chain enhancer, and the modifiedmurine dhfr cDNA for selection by methotrexate (MTX).

The heavy chain cassette was transferred accordingly into an expressionvector encoding the human IgG1 gene, the immunoglobulin heavy chainenhancer, and the neomycin resistance gene for selection.

Both heavy and light chain are in a configuration in the expressionvectors that resembles the genomic configuration of rearrangedimmunoglobulin genes which is thought to be crucial for high levelexpression.

For Antibody production the above vectors are co-transfected into anappropriate host cell line, e.g. the SP2/0 cell line, cells containingthe vector sequences are selected by methotrexate selection, andselected cell lines are cultured to express the AAL160 antibody.Alternatively a GS based amplification/selection system such as thatdescribed in EP 0256055 B, EP 0323997 B or European patent application89303964.4 may be used, in which case the dhfr selectable marker isreplaced by a GS coding sequence.

Example 2 Biochemical and Biological Data

The monoclonal antibody AAL160 is found to neutralize the activity ofinterleukin-1β in vitro. The monoclonal antibody is furthercharacterized for its binding to recombinant human IL-1β Biacoreanalysis. The mode of neutralization is assessed by competitive bindingstudies with soluble IL-1 receptors. The biological activity of theantibody AAL 160 towards recombinant and naturally produced IL-1β isdetermined in primary human cells (Example 3), responsive to stimulationby IL-1β.

2.1 Determination of Dissociation Equilibrium Constant

The association and dissociation rate constant for the binding ofrecombinant human IL-1β to AAL160 is determined by BIAcore analysis.AAL160 is immobilized, and binding of recombinant IL-1β in aconcentration range from 0.5 to 12 nM is measured by surface plasmonresonance. The chosen format permits treating the binding event of IL-1βto AAL160 according to a 1:1 stoichiometry. Data analysis is performedusing the BIAevaluation software.

Association rate (n = 15) (3.91 ± 0.14) × 10⁵   Mean ± SEM constant[M⁻¹s⁻¹] Dissociation rate (n = 15) (1.53 ± 0.05) × 10⁻⁴ Mean ± SEMconstant [s⁻¹] Dissociation (n = 15) (396.6 ± 19.5) × 10⁻¹²   Mean ± SEMequilibrium constant K_(D) [M]

AAL160 binds to recombinant human IL-1β with a high affinity.

2.2. Competitive Inhibition of Binding to Soluble IL-1 Receptors

Binding Competition Study with Soluble IL-1 Type I and II Receptors

Competition between AAL160 and soluble human IL-1 type I and type IIreceptors is measured by Biacore. AAL160 is immobilized on the chipsurface and recombinant human IL-1β (8 nM) is injected for binding toAAL160 in absence or presence of increasing concentrations ofrecombinant human soluble receptor I (0-10 nM) or receptor II (0-80 nM).The results obtained are shown in FIG. 1. Binding of NVP AAL 160 NX-1 toIL-1β is competitive with both IL-1 receptor type I and type II

2.3. Reactivity Profile to Human IL-1α, Human IL-1Ra, and IL-1β fromRodent and Monkey Species

The reactivity profile of AAL160 to human IL-1α, IL-1RA, and murine,rat, rabbit and cynomolgus monkey IL-1β is determined by Biacoreanalysis. AAL160 is immobilized, and the cytokines examined are appliedat a concentration of 8 nM (or 20 nM in the case of IL-1β.)

Percent of total binding ± SEM Human IL-1β 100 Human IL-1α 0.7 ± 0.7 (n= 3) Human IL-1Ra 1.2 ± 1.2 (n = 3) Mouse IL-1β 2.8 ± 1.5 (n = 3) RatIL-1β 3.0 ± 2.5 (n = 3) Cynomolgus IL-1β 96.4 ± 6.8 (n = 3)  RabbitIL-1β 12.1 ± 2.3 (n = 4) AAL160 does not significantly crossreact with human IL-1α, human IL-1Ra,or murine, rat or rabbit IL-1β. The reactivity towards cynomolgus monkeyIL-1β is virtually identical to the human cytokine.

Example 3 Neutralization of IL-β-Dependent Production of PGE₂ andInterleukin-6 by Primary Human Fibroblasts

The production of PGE₂ and IL-6 in primary human dermal fibroblasts isdependent on IL-1β. TNF-α alone cannot efficiently induce theseinflammatory mediators, but synergizes with IL-1. Primary dermalfibroblasts are used as a surrogate model for IL-1 induced cellularactivation.

Primary human fibroblasts are stimulated with recombinant IL-1β orconditioned medium obtained from LPS-stimulated human PBMCs in thepresence of various concentrations of AAL160 or IL-1RA ranging from 6 to18,000 pM. The chimeric anti-CD25 antibody SIMULECT® (basiliximab) isused as a matched isotype control. Supernatant is taken 16 hours afterstimulation and is assayed for IL-6 by ELISA or PGE₂ by RIA.

AAL160 IL-1 Ra IC₅₀ ± SEM (n ≧ 3) IC₅₀ ± SEM (n ≧ 3) IL-6 secretion 0.34± 0.037 nM n.d. Recombinant IL-6 secretion 0.6 ± 0.09 nM 0.03 ± 0.001 nMcond. Medium PGE₂ production 0.79 ± 0.17 nM  n.d. cond. MediumAAL160 effectively blocks production of IL-6 and PGE₂ in human dermalfibroblasts with an IC₅₀ similar for both the recombinant and naturalIL-1β.

Example 4 In Vivo Efficacy and Duration of Action of AAL160 Efficacy:

The in vivo efficacy of the anti-huIL-1β antibody, AAL 160 is tested ina rat model where fever is induced by an i.v. injection of huIL-1β (100ng/rat). The antibody causes a dose related inhibition of the feverresponse over the dose range 1, 3 and 10 μg/kg i.v. (n=6 rats)—see FIG.2. CHI 621 (SIMULECT®, basiliximab) is used as the control antibody.

Duration of Action:

The duration of action of AAL-160 is investigated in rat IL-1β-inducedfever as follows: The antibody is injected i.v. either 24 hours or 30minutes (standard protocol) before the induction of fever by an i.v.injection of human IL-1β and body temperature measured 2 and 4 hourslater. A similar degree of inhibition of the fever response is seen atboth times (see FIG. 3). As expected, the control antibody CHI 621(SIMULECT®, basiliximab) is ineffective at both time-points. Thisfinding indicates that the AAL160 human antibody is present in an activeform for at least 24 hours in the rat and is not metabolised, excretedor bound in the tissues during this time.

Example 5 X-Ray Studies of AAL160 Fab and its Complex with IL-1βStructure Determination of AAL160 Fab at 2.0□ Resolution:

A 2.0□ resolution data set of very good quality (R_(sym)=0.051,completeness=99.9%, redundancy=8.2) was collected from an Fab crystalgrown by the vapour diffusion in hanging drop technique, at pH 9.5 in50% PEG 200, 0.1 M CHES. The crystal was in space group P2₁2₁2₁ withunit cell dimensions a=62.17 Å b=89.83 Å c=123.73 Å and one Fab moleculeper symmetric unit (Matthews coefficient: 3.6 Å³/Da, estimated solventcontent: 66%). The structure was determined by molecular replacement andwas refined to a final crystallographic R-factor of 0.209 (freeR-factor=0.261). The final model includes residues 1-213 of the lightchain, 1-131 and 138-218 of the heavy chain, 387 water molecules and 1PEG molecule. The final electron density is well defined for all CDRresidues but Trp 94 (CDR3) of the light chain. The position of theside-chain of this residue is ill defined in the two crystal forms whichwere examined to date, thus suggesting that it is highly mobile in theabsence of a bound antigen.

Crystallization of the Fab complex with IL-1β and preliminaryexperimental model of the complex: a few crystals of AAL160 Fab incomplex with the antigen IL-1β were obtained from a 76 mg ml stocksolution of the 1:1 complex in 2.0M ammonium sulfate, 0.1M Tris pH 8.5.The crystals grew very slowly over a period of several weeks. Theydiffracted weakly to about 3.2□ on the home source. A preliminary dataset was collected and molecular replacement was attempted using the highresolution structures of the free Fab and of human IL-1β (J. P. Priestleet al., EMBO J. 7, 339 (1988)) as starting models. The calculationsyielded a very clear and unambiguous solution when the Fv and Fc partsof the Fab were used as separate modules (correlation 67.1%, R-factor0.354 after the AMORE FITTING step, using data between 8.0 and 3.5□).The subsequent comparison of the free and bound forms of the Fab showedthat the elbow angle is very different in the two structures. Theresults of the molecular replacement calculations provide a firstmolecular model of the interactions between the antigen IL-1β and themonoclonal antibody AAL160. A preliminary analysis of these interactionsindicates that 1) IL-1β makes tight interactions to all three CDRs ofthe heavy chain and to CDR3 of the light chain. In contrast, fewinteractions if any involve CDR1 and CDR of the light chain. 2) The loopcomprising residues Gly 22, Pro 23, Tyr 24 and Glu 25 of mature IL-1βbinds at the centre of the antigen-combining site and thus appears to bea key component of the epitope. Interestingly enough, this loop is notlocated in the region of the molecule that differs most from mouseIL-1β. Pro 23, Tyr 24 and Glu 25 are conserved, but residue 22 is a Glyin human IL-1β and an Asp in mouse IL-1β. Comparison of the crystalstructures of human (PDB entry 2i1b) and mouse IL-1β (PDB entry 8i1b)shows that this point mutation results in a very different conformationof the main-chain around Pro 23. This local structural difference isconsistent with the observed lack of cross-reactivity of AAL160 withrespect to the mouse cytokine.

1-12. (canceled)
 13. A polynucleotide construct that encodes animmunoglobulin heavy chain or fragment thereof of an IL-1β bindingmolecule, wherein said immunoglobulin heavy chain comprises a heavychain constant domain or fragment thereof and a heavy chain variabledomain or fragment thereof, and wherein said heavy chain variable domaincomprises framework regions and a first region consisting of amino acids31 to 35 of SEQ ID NO:1, a second region consisting of amino acids 50 to66 of SEQ ID NO:1, and a third region consisting of amino acids 99 to107 of SEQ ID NO:1.
 14. A polynucleotide construct that encodes animmunoglobulin light chain or fragment thereof IL-1β binding molecule,wherein said immunoglobulin light chain comprises a light chain constantdomain or fragment thereof and a light chain variable domain or fragmentthereof, and wherein said light chain variable domain comprisesframework regions and a first region consisting of amino acids 24 to 34of SEQ ID NO:2, a second region consisting of amino acids 50 to 56 ofSEQ ID NO:2 and a third region consisting of amino acids 89 to 97 of SEQID NO:2.
 15. A polynucleotide encoding an isolated IL-1β bindingmolecule comprising a heavy chain variable domain comprising SEQ ID NO:1or a light chain variable domain comprising SEQ ID NO:2.
 16. Apolynucleotide encoding an isolated IL-1β binding molecule comprisingthe three CDRs of SEQ ID NO:1 or the three CDRs of SEQ ID NO:2.
 17. Apolynucleotide encoding an isolated IL-1β binding molecule comprisingSEQ ID NO:5. SEQ ID NO:6 and SEQ ID NO:7 or SEQ ID NO:8, SEQ ID NO:9 andSEQ ID NO:10.
 18. An expression vector able to replicate in aprokaryotic or eukaryotic cell which comprises the construct accordingto claim
 13. 19. An expression vector able to replicate in a prokaryoticor eukaryotic cell which comprises the construct according to claim 14.20. An expression vector able to replicate in a prokaryotic oreukaryotic cell which comprises the polynucleotide according to claim15.
 21. An expression vector able to replicate in a prokaryotic oreukaryotic cell which comprises the polynucleotide according to claim16.
 22. An expression vector able to replicate in a prokaryotic oreukaryotic cell which comprises the polynucleotide according to claim17.
 23. A prokaryotic or eukaryotic host cell comprising the constructaccording to claim
 13. 24. A prokaryotic or eukaryotic host cellcomprising the construct according to claim
 14. 25. A prokaryotic oreukaryotic host cell comprising the polynucleotide according to claim15.
 26. A prokaryotic or eukaryotic host cell comprising thepolynucleotide according to claim
 16. 27. A prokaryotic or eukaryotichost cell comprising the polynucleotide according to claim
 17. 28. Aprocess for the production of an IL-1β binding molecule which comprises(i) culturing the host cell of claim 23 to yield a cell culture and (ii)recovering the IL-1β binding molecule from said cell culture.
 29. Aprocess for the production of an IL-1β binding molecule which comprises(i) culturing the host cell of claim 24 to yield a cell culture and (ii)recovering the IL-1β binding molecule from said cell culture.
 30. Aprocess for the production of an IL-1β binding molecule which comprises(i) culturing the host cell of claim 25 to yield a cell culture and (ii)recovering the IL-1β binding molecule from said cell culture.
 31. Aprocess for the production of an IL-1β binding molecule which comprises(i) culturing the host cell of claim 26 to yield a cell culture and (ii)recovering the IL-1β binding molecule from said cell culture.
 32. Aprocess for the production of an IL-1β binding molecule which comprises(i) culturing the host cell of claim 27 to yield a cell culture and (ii)recovering the IL-1β binding molecule from said cell culture.
 33. Amethod for treating an IL-1β mediated disease or disorder or aninflammatory disease or disorder in a subject in need thereof comprisingadministering to the subject an effective amount of an IL-1β bindingmolecule comprising a heavy chain variable domain comprising SEQ IDNO:1.
 34. A method for treating an IL-1β mediated disease or disorder oran inflammatory disease or disorder in a subject in need thereofcomprising administering to the subject an effective amount of an IL-1βbinding molecule comprising a light chain variable domain comprising SEQID NO:2.
 35. The method of claim 33, wherein the IL-1β binding moleculefurther comprises a light chain variable domain comprising SEQ ID NO:2.36. The method of claim 34, wherein the IL-1β binding molecule furthercomprises a heavy chain variable domain comprising SEQ ID NO:1.
 37. Amethod for treating an IL-1β mediated disease or disorder or aninflammatory disease or disorder in a subject in need thereof comprisingadministering to the subject an effective amount of an isolated IL-1βbinding molecule comprising a heavy chain variable domain comprising SEQID NO:1 and a light chain variable domain comprising SEQ ID NO:2.
 38. Amethod for treating an IL-1β mediated disease or disorder or aninflammatory disease or disorder in a subject in need thereof comprisingadministering to the subject an effective amount of an isolated IL-1βbinding molecule comprising a heavy chain variable domain comprising thethree CDRs of SEQ ID NO:1 and a light chain variable domain comprisingthe three CDRs of SEQ ID NO:2. 39-40. (canceled)
 41. A method fortreating an IL-1β mediated disease or disorder or an inflammatorydisease or disorder in a subject in need thereof comprisingadministering to the subject an effective amount of an isolated IL-1βbinding molecule that is capable of inhibiting the binding of IL-1β tothe IL-1β receptor to the same extent as an IL-1β binding moleculecomprising a heavy chain variable domain comprising SEQ ID NO:1 and alight chain variable domain comprising SEQ ID NO:2.
 42. A method fortreating an IL-1β mediated disease or disorder or an inflammatorydisease or disorder in a subject in need thereof comprisingadministering to the subject an effective amount of an isolated IL-1βbinding molecule that is capable of inhibiting the binding of IL-1β tothe IL-1β receptor to the same extent as an IL-1β binding moleculecomprising a heavy chain variable domain comprising the three CDRs ofSEQ ID NO:1 and a light chain variable domain comprising the three CDRsof SEQ ID NO:2.
 43. The method according to claim 38, wherein said IL-1βbinding molecule is a human antibody.
 44. A method for treating an IL-1βmediated disease or disorder or an inflammatory disease or disorder in asubject in need thereof comprising administering to the subject aneffective amount of an IL-1β binding molecule encoded by thepolynucleotide of claim
 15. 45. A method for treating an IL-1β mediateddisease or disorder or an inflammatory disease or disorder in a subjectin need thereof comprising administering to the subject an effectiveamount of an IL-1β binding molecule encoded by the polynucleotide ofclaim
 16. 46. A method for treating an IL-1β mediated disease ordisorder or an inflammatory disease or disorder in a subject in needthereof comprising administering to the subject an effective amount ofan IL-1β binding molecule encoded by the polynucleotide of claim
 17. 47.The method according to claim 46, wherein said IL-1β binding molecule isa human antibody.
 48. A method for modulating IL-1β in a subject,comprising administering to the subject an isolated IL-1β bindingmolecule encoded by the polynucleotide of claim
 15. 49. A method formodulating IL-1β in a subject, comprising administering to the subjectan isolated IL-1β binding molecule encoded by the polynucleotide ofclaim
 16. 50. A method for modulating IL-1β in a subject, comprisingadministering to the subject an isolated IL-1β binding molecule encodedby the polynucleotide of claim
 17. 51. A method for binding IL-1β in asubject, comprising administering to the subject an isolated IL-1βbinding molecule encoded by the polynucleotide of claim
 15. 52. A methodfor binding IL-1β in a subject, comprising administering to the subjectan isolated IL-1β binding molecule encoded by the polynucleotide ofclaim
 16. 53. A method for binding IL-1β in a subject, comprisingadministering to the subject an isolated IL-1β binding molecule encodedby the polynucleotide of claim
 17. 54. A method for administering anisolated IL-1β binding molecule to a subject, comprising administeringto the subject a pharmaceutical composition comprising an isolated IL-1βbinding molecule encoded by the polynucleotide of claim
 15. 55. A methodfor administering an isolated IL-1β binding molecule to a subject,comprising administering to the subject a pharmaceutical compositioncomprising an isolated IL-1β binding molecule encoded by thepolynucleotide of claim
 16. 56. A method for administering an isolatedIL-1β binding molecule to a subject, comprising administering to thesubject a pharmaceutical composition comprising an isolated IL-1βbinding molecule encoded by the polynucleotide of claim
 17. 57. Anisolated IL-1β binding molecule that is capable of inhibiting thebinding of IL-1β to the IL-1β receptor to the same extent as an IL-1βbinding molecule encoded by the polynucleotide of claim
 15. 58. Anisolated IL-1β binding molecule that is capable of inhibiting thebinding of IL-1β to the IL-1β receptor to the same extent as an IL-1βbinding molecule encoded by the polynucleotide of claim
 16. 59. Anisolated IL-1β binding molecule that is capable of inhibiting thebinding of IL-1β to the IL-1β receptor to the same extent as an IL-1βbinding molecule encoded by the polynucleotide of claim 17.